miScript miRNA Mimics

合成miRNAを用いたmiRNA機能および遺伝子制御に関する研究

  • 内在性mature miRNAと同様の機能を持つ即トランスフェクト可能なMimic
  • miRBase に登録されているヒト、マウス、ラットmiRNA に対応
  • 様々なフォーマット、スケール、グレードで提供
  • miRBaseに登録されていないmiRNAでもカスタムデザインで対応可能
  • GeneGlobeから簡単に注文

miScript miRNA Mimics は、細胞へのトランスフェクション後に内在性のmature miRNA と同様の機能を示す合成二本鎖RNA です。miScript miRNA Mimics は、細胞培養グレード(純度90%以上)あるいは動物グレード(HPLC 精製、in vivo アプリケーション用)でお届けしています。miScript miRNA Mimic Platesではデザイン済みあるいはカスタム合成のmiScript miRNA Mimicsを、96 ウェルあるいは384 ウェルプレートでお届けしています。同類の実験用にmiScript miRNA Inhibitorsもお使いいただけます。miRNA Mimic トランスフェクションは HiPerFect Transfection Reagent、miRNA Mimic とDNA のコトランスフェクションはAttractene Transfection Reagent をご利用ください。

Search in GeneGlobe
You can search for the following terms:
  • Entrez Gene IDs (e.g., 835)
  • RefSeq IDs (e.g., NM_032983, NP_116765)
  • Gene symbols (e.g., CASP2)
  • Cat. no. (e.g., SI00299551, QT01342509)
  • Sanger ID or Accession (e.g., hsa-let-7b, MI0000063)
  • CpG loci identification numbers (CG#) (e.g., CG17753661)

Do not enter species information in the Search box. Use the drop-down list to select your species of interest.
When searching for miRNAs, do not omit the hyphens. Use hyphens or spaces (e.g., search for hsa-let-7b or hsa let 7b. Do not search for hsalet7b).

Advanced search settings



Species:
Products

Product Product no. Cat. no. List price:
 
 
Show details
    varies
Can't order online?
To place an order via phone, email or for requesting a quote, please provide the product’s name, number and catalog number.
Product Cat. no. List price:
miScript miRNA Mimic
1 nmol (cell-culture grade), 5 nmol (cell-culture grade), or 20 nmol (animal grade) miScript miRNA Mimic, provided in tubes
Varies
Details

Can’t order online or need plate formats? Order Forms and Plate Orders

miScript miRNA Mimics are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.


  • Main Image Navi
Comparable downregulation by endogenous miRNA and miScript miRNA Mimic.|miScript miRNA Mimic stable for 72 hours.|Downregulation at protein level after miR-1 mimic transfection.|
miR-1 is not endogenously expressed in HeLa S3 cells. miR-16 is endogenously expressed at high levels in HeLa S3 cells (Lagos-Quintana, M. et al. [2001] Identification of novel genes coding for small expressed RNAs. Science 294, 853.). Using HiPerFect Transfection Reagent, HeLa S3 cells (6 x 104 cells/well) were transfected with a luciferase reporter construct with a binding site for miR-1 in the 3' UTR; cotransfected with a luciferase reporter construct with a binding site for miR-1 in the 3' UTR and with a miR-1 Mimic; or transfected with a luciferase reporter construct with a binding site for miR-16 in the 3' UTR. Twenty-four hours later, luciferase activity was measured. miR-1 Mimic resulted in a similar decrease in luciferase level as endogenous miR-16.|HeLa S3 cells (6 x 104 cells/well) were transfected with a negative control or an miR-1 Mimic using HiPerFect Transfection Reagent. Twenty-four hours prior to measurement, cells were transfected with a luciferase reporter construct carrying an miR-1 binding site using Attractene Transfection Reagent. After the time points indicated, miR-1 levels were detected by measuring luciferase activity.|HDAC4 is predicted to be a target of miR-1 and has 2 miR-1 binding sites in the 3' UTR (Chen, J.F. et al. [2006] The role of microRNA-1 and microRNA-133 in skeletal muscle proliferation and differentiation. Nat. Genet. 38, 228.). HeLa S3 cells (8 x 104 cells/well) were transfected using HiPerFect Transfection Reagent and a range of concentrations of miR-1 Mimic. miR-1 #1, miR-1 #2, and miR-1 #3 are different batches of miR-1 Mimic. After 24 and 48 hours, [A] HDAC4 protein level was analyzed by western blotting. Tubulin was also analyzed as an internal control (not shown). No effect was observed after 24 hours. After 48 hours, protein level was significantly decreased. [B] HDAC4 transcript level was analyzed by real-time PCR using the miScript System after transfection using 50 nM miR-1 Mimic. The negative control was AllStars Negative Control. Results were normalized to GAPDH and to the negative control and are presented as 2-ΔΔCT. Transcript levels did not change significantly at either the 24-hour or 48-hour time point.|
Performance
miScript miRNA mimics は内在性miRNAで観察される現象と同様に遺伝子発現抑制を引き起こします(図 “内在性miRNA とmiScript miRNA mimicで同等の発現抑制を実現”)。観察された遺伝子発現抑制は翻訳レベルで行なわれます(図“miR-1 Mimic のトランスフェクション後のタンパク質抑制効果”)。miScript miRNA mimics はトランスフェクション72 時間後まで遺伝子発現を効果的に阻害します(図 “72 時間安定なmiScript miRNA mimic”)。
Principe

Mimicsのトランスフェクションおよびその後の遺伝子発現解析や表現型の解析は特定のmiRNA の標的遺伝子の同定や機能を解明するために有効です。これらの実験により、個別のmiRNAが標的とする遺伝子の確認や個々のmiRNA間違った制御による生物学的影響を調べることができます。Mimics のトランスフェクション後に遺伝子発現が低下すれば、研究対象のmiRNA がその遺伝子の制御に関与している証拠になります。miRNA の作用を阻害する miScript miRNA Inhibitors も同様の研究目的で使用できます。一般に行なう実験では、Mimicあるいは Inhibitorのトランスフェクション、あるいはレポーター遺伝子を融合したmiRNA結合部位を持つベクターコンストラクトとのコトランスフェクションを行います。ダウンストリーム解析としてはレポーターアッセイ、リアルタイムPCR、マイクロアレイ解析、タンパク質解析があります。

Procédure

現在の miRBase (microRNAのデータベース)に登録済みのヒト、マウス、ラットmiRNA に対応したmiScript miRNA mimicsが入手できます。最新miRBaseバージョンに対応したMimic およびInhibitor miRNA をデザインするために、QIAGEN のGeneGlobe データベースは常にアップデートされています。従って入手できる最新かつ正確なmiRNA配列情報で常に実験を行うことができます。目的のmiRNA がmiRBase に登録されていない場合には、お客様のヒト、マウス、ラットmiRNA に対応するmimics をカスタムデザインできます。miScript miRNA 製品の カスタムデザインページ で、入手したいmimic の名称をつけ、ターゲット配列を入力したのち、“Create”ボタンを押すだけで、目的のカスタムデザインmimic が簡単に入手できます。

様々なフォーマットを取り揃えているので、少数のMimic を用いたロースループット解析でも数百のMimic を用いたハイスループットスクリーニングでも可能です。miScript miRNA mimics は、培養細胞に最適なグレード (独自の合成、精製プロセスにより90% 以上の純度) のmiRNAをチューブ(1 nmolか5 nmol)でお届けします。in vivo 用にはHPLC 精製した動物実験に最適なグレードのmiScript miRNA mimics を20 nmol の収量で注文できます(チューブ)。ミドル~ハイスループットなアプリケーション用には、実験の条件に応じて96ウェルプレート(1 nmol か5 nmol)、あるいは384ウェルプレート(1 nmol)を提供しています。ご希望のレイアウトでデザイン済み(miRBaseに掲載)あるいはカスタムデザイン(miRBaseに未掲載) miScript miRNA mimicsの エクセルファイルをアップロードすれば、プレートの確認と注文をしていただくことができます。

Applications
miRNA 研究
遺伝子制御研究
Feature
Specifications
Design Predesigned and custom design avaliable
Format Single tubes, 96-well & 384 well plates
Modification None
Scale 1 nmol, 5 nmol, 20 nmol
Sequence provided No
Species Human, mouse, rat

You are not authorized to download the resource

Safety Data Sheets
1
Images
Comparable Downregulation by Endogenous miRNA and miScript miRNA Mimic Comparable Downregulation by Endogenous miRNA and miScript miRNA Mimic
Comparable downregulation by endogenous miRNA and miScript miRNA Mimic.
miR-1 is not endogenously expressed in HeLa S3 cells. miR-16 is endogenously expressed at high levels in HeLa S3 cells (Lagos-Quintana, M. et al. [2001] Identification of novel genes coding for small expressed RNAs. Science 294, 853.). Using HiPerFect Transfection Reagent, HeLa S3 cells (6 x 104 cells/well) were transfected with a luciferase reporter construct with a binding site for miR-1 in the 3' UTR; cotransfected with a luciferase reporter construct with a binding site for miR-1 in the 3' UTR and with a miR-1 Mimic; or transfected with a luciferase reporter construct with a binding site for miR-16 in the 3' UTR. Twenty-four hours later, luciferase activity was measured. miR-1 Mimic resulted in a similar decrease in luciferase level as endogenous miR-16.
miScript miRNA Mimic Stable for 72 Hours
miScript miRNA Mimic stable for 72 hours.
HeLa S3 cells (6 x 104 cells/well) were transfected with a negative control or an miR-1 Mimic using HiPerFect Transfection Reagent. Twenty-four hours prior to measurement, cells were transfected with a luciferase reporter construct carrying an miR-1 binding site using Attractene Transfection Reagent. After the time points indicated, miR-1 levels were detected by measuring luciferase activity.
Downregulation at Protein Level after miR-1 Mimic Transfection
Downregulation at protein level after miR-1 mimic transfection.
HDAC4 is predicted to be a target of miR-1 and has 2 miR-1 binding sites in the 3' UTR (Chen, J.F. et al. [2006] The role of microRNA-1 and microRNA-133 in skeletal muscle proliferation and differentiation. Nat. Genet. 38, 228.). HeLa S3 cells (8 x 104 cells/well) were transfected using HiPerFect Transfection Reagent and a range of concentrations of miR-1 Mimic. miR-1 #1, miR-1 #2, and miR-1 #3 are different batches of miR-1 Mimic. After 24 and 48 hours, [A] HDAC4 protein level was analyzed by western blotting. Tubulin was also analyzed as an internal control (not shown). No effect was observed after 24 hours. After 48 hours, protein level was significantly decreased. [B] HDAC4 transcript level was analyzed by real-time PCR using the miScript System after transfection using 50 nM miR-1 Mimic. The negative control was AllStars Negative Control. Results were normalized to GAPDH and to the negative control and are presented as 2-ΔΔCT. Transcript levels did not change significantly at either the 24-hour or 48-hour time point.